Fluorescent probes for the detection of reactive oxygen species in human spermatozoa
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10316/105804 https://doi.org/10.4103/aja.aja_132_19 |
Resumo: | Reactive oxygen species (ROS) production is a by-product of mitochondrial activity and is necessary for the acquisition of the capacitated state, a requirement for functional spermatozoa. However, an increase in oxidative stress, due to an abnormal production of ROS, has been shown to be related to loss of sperm function, highlighting the importance of an accurate detection of sperm ROS, given the specific nature of this cell. In this work, we tested a variety of commercially available fluorescent probes to detect ROS and reactive nitrogen species (RNS) in human sperm, to define their specificity. Using both flow cytometry (FC) and fluorescence microscopy (FM), we confirmed that MitoSOX™ Red and dihydroethidium (DHE) detect superoxide anion (as determined using antimycin A as a positive control), while DAF-2A detects reactive nitrogen species (namely, nitric oxide). For the first time, we also report that RedoxSensor™ Red CC-1, CellROX® Orange Reagent, and MitoPY1 seem to be mostly sensitive to hydrogen peroxide, but not superoxide. Furthermore, mean fluorescence intensity (and not percentage of labeled cells) is the main parameter that can be reproducibly monitored using this type of methodology. |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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7160 |
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Fluorescent probes for the detection of reactive oxygen species in human spermatozoaflow cytometryfluorescent probeshuman spermatozoaoxidative stressreactive oxygen speciesEthidiumFlow CytometryHumansHydrogen PeroxideMaleMicroscopy, FluorescenceOrganophosphorus CompoundsPhenanthridinesPiperazinesReactive Nitrogen SpeciesReactive Oxygen SpeciesSpermatozoaSpiro CompoundsSuperoxidesFluorescent DyesReactive oxygen species (ROS) production is a by-product of mitochondrial activity and is necessary for the acquisition of the capacitated state, a requirement for functional spermatozoa. However, an increase in oxidative stress, due to an abnormal production of ROS, has been shown to be related to loss of sperm function, highlighting the importance of an accurate detection of sperm ROS, given the specific nature of this cell. In this work, we tested a variety of commercially available fluorescent probes to detect ROS and reactive nitrogen species (RNS) in human sperm, to define their specificity. Using both flow cytometry (FC) and fluorescence microscopy (FM), we confirmed that MitoSOX™ Red and dihydroethidium (DHE) detect superoxide anion (as determined using antimycin A as a positive control), while DAF-2A detects reactive nitrogen species (namely, nitric oxide). For the first time, we also report that RedoxSensor™ Red CC-1, CellROX® Orange Reagent, and MitoPY1 seem to be mostly sensitive to hydrogen peroxide, but not superoxide. Furthermore, mean fluorescence intensity (and not percentage of labeled cells) is the main parameter that can be reproducibly monitored using this type of methodology.Wolters Kluwer Health2020info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://hdl.handle.net/10316/105804http://hdl.handle.net/10316/105804https://doi.org/10.4103/aja.aja_132_19eng1008-682XEscada-Rebelo, SaraMora, FranciscaSousa, AnaAlmeida Santos, TeresaPaiva, ArturRamalho-Santos, Joãoinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-03-08T21:31:10Zoai:estudogeral.uc.pt:10316/105804Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T21:22:18.341652Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Fluorescent probes for the detection of reactive oxygen species in human spermatozoa |
title |
Fluorescent probes for the detection of reactive oxygen species in human spermatozoa |
spellingShingle |
Fluorescent probes for the detection of reactive oxygen species in human spermatozoa Escada-Rebelo, Sara flow cytometry fluorescent probes human spermatozoa oxidative stress reactive oxygen species Ethidium Flow Cytometry Humans Hydrogen Peroxide Male Microscopy, Fluorescence Organophosphorus Compounds Phenanthridines Piperazines Reactive Nitrogen Species Reactive Oxygen Species Spermatozoa Spiro Compounds Superoxides Fluorescent Dyes |
title_short |
Fluorescent probes for the detection of reactive oxygen species in human spermatozoa |
title_full |
Fluorescent probes for the detection of reactive oxygen species in human spermatozoa |
title_fullStr |
Fluorescent probes for the detection of reactive oxygen species in human spermatozoa |
title_full_unstemmed |
Fluorescent probes for the detection of reactive oxygen species in human spermatozoa |
title_sort |
Fluorescent probes for the detection of reactive oxygen species in human spermatozoa |
author |
Escada-Rebelo, Sara |
author_facet |
Escada-Rebelo, Sara Mora, Francisca Sousa, Ana Almeida Santos, Teresa Paiva, Artur Ramalho-Santos, João |
author_role |
author |
author2 |
Mora, Francisca Sousa, Ana Almeida Santos, Teresa Paiva, Artur Ramalho-Santos, João |
author2_role |
author author author author author |
dc.contributor.author.fl_str_mv |
Escada-Rebelo, Sara Mora, Francisca Sousa, Ana Almeida Santos, Teresa Paiva, Artur Ramalho-Santos, João |
dc.subject.por.fl_str_mv |
flow cytometry fluorescent probes human spermatozoa oxidative stress reactive oxygen species Ethidium Flow Cytometry Humans Hydrogen Peroxide Male Microscopy, Fluorescence Organophosphorus Compounds Phenanthridines Piperazines Reactive Nitrogen Species Reactive Oxygen Species Spermatozoa Spiro Compounds Superoxides Fluorescent Dyes |
topic |
flow cytometry fluorescent probes human spermatozoa oxidative stress reactive oxygen species Ethidium Flow Cytometry Humans Hydrogen Peroxide Male Microscopy, Fluorescence Organophosphorus Compounds Phenanthridines Piperazines Reactive Nitrogen Species Reactive Oxygen Species Spermatozoa Spiro Compounds Superoxides Fluorescent Dyes |
description |
Reactive oxygen species (ROS) production is a by-product of mitochondrial activity and is necessary for the acquisition of the capacitated state, a requirement for functional spermatozoa. However, an increase in oxidative stress, due to an abnormal production of ROS, has been shown to be related to loss of sperm function, highlighting the importance of an accurate detection of sperm ROS, given the specific nature of this cell. In this work, we tested a variety of commercially available fluorescent probes to detect ROS and reactive nitrogen species (RNS) in human sperm, to define their specificity. Using both flow cytometry (FC) and fluorescence microscopy (FM), we confirmed that MitoSOX™ Red and dihydroethidium (DHE) detect superoxide anion (as determined using antimycin A as a positive control), while DAF-2A detects reactive nitrogen species (namely, nitric oxide). For the first time, we also report that RedoxSensor™ Red CC-1, CellROX® Orange Reagent, and MitoPY1 seem to be mostly sensitive to hydrogen peroxide, but not superoxide. Furthermore, mean fluorescence intensity (and not percentage of labeled cells) is the main parameter that can be reproducibly monitored using this type of methodology. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10316/105804 http://hdl.handle.net/10316/105804 https://doi.org/10.4103/aja.aja_132_19 |
url |
http://hdl.handle.net/10316/105804 https://doi.org/10.4103/aja.aja_132_19 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
1008-682X |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Wolters Kluwer Health |
publisher.none.fl_str_mv |
Wolters Kluwer Health |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
instname_str |
Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
instacron_str |
RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
repository.mail.fl_str_mv |
|
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1799134112751026176 |